Pyrrolotriazines as alk inhibitors

ABSTRACT

This application provides compounds of the general formula (I) 
     
       
         
         
             
             
         
       
     
     and/or a salt thereof, where X, R 1  and R 2  are as defined herein. Compositions and therapeutic uses are also described.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.14/944,668, filed Nov. 18, 2015, which is a continuation ofInternational Application No. PCT/US2014/039755, filed May 28, 2014,which claims the benefit of U.S. provisional application No. 61/828,451,filed May 29, 2013, the disclosures of which are incorporated herein byreference in their entireties.

BACKGROUND

Anaplastic Lymphoma Kinase (ALK) is a cell membrane-spanning receptortyrosine kinase, which belongs to the insulin receptor subfamily. Themost abundant expression of ALK occurs in the neonatal brain, suggestinga possible role for ALK in brain development (Duyster, J. et al.,Oncogene, 2001, 20, 5623-5637).

ALK is also implicated in the progression of certain tumors. Forexample, approximately sixty percent of anaplastic large cell lymphomas(ALCL) are associated with a chromosome mutation that generates a fusionprotein consisting of nucleophosmin (NPM) and the intracellular domainof ALK. (Armitage, J. O. et al., Cancer: Principle and Practice ofOncology, 6^(th) edition, 2001, 2256-2316; Kutok J. L. & Aster J. C., J.Clin. Oncol., 2002, 20, 3691-3702). This mutant protein, NPM-ALK,possesses a constitutively active tyrosine kinase domain that isresponsible for its oncogenic property through activation of downstreameffectors. (Falini, B. et al., Blood, 1999, 94, 3509-3515; Morris, S. W.et al., Brit. J. Haematol., 2001, 113, 275-295; Duyster et al.; Kutok &Aster). In addition, the transforming EML4ALK fusion gene has beenidentified in non-small-cell lung cancer (NSCLC) patients (Soda, M., etal., Nature, 2007, 448, 561-566) and represents another in a list of ALKfusion proteins that are promising targets for ALK inhibitor therapy.Experimental data have demonstrated that the aberrant expression ofconstitutively active ALK is directly implicated in the pathogenesis ofALCL and that inhibition of ALK can markedly impair the growth of ALK+lymphoma cells (Kuefer, Mu et al. Blood, 1997, 90, 2901-2910; Bai, R. Y.et al., Mol. Cell Biol., 1998, 18, 6951-6961; Bai, R. Y. et al., Blood,2000, 96, 4319-4327; Ergin, M. et al., Exp. Hematol., 2001, 29,1082-1090; Slupianek, A. et al., Cancer Res., 2001, 61, 2194-2199;Turturro, F. et al., Clin. Cancer Res., 2002, 8, 240-245). Theconstitutively activated chimeric ALK has also been demonstrated inabout 60% of inflammatory myofibroblastic tumors (IMTs), a slow-growingsarcoma that mainly affects children and young adults. (Lawrence, B. etal., Am. 1 Pathol., 2000, 157, 377-384; Duyster et al.).

In addition, ALK and its putative ligand, pleiotrophin, areoverexpressed in human glioblastomas (Stoica, G. et al., J. Biol. Chem.,2001, 276, 16772-16779). In mouse studies, depletion of ALK reducedglioblastoma tumor growth and prolonged animal survival (Powers, C. etal., J. Biol. Chem., 2002, 277, 14153-14158; Mentlein, R. et al, J.Neurochem., 2002, 83, 747-753).

It is thought that ALK inhibitors would permit durable cures whencombined with current chemotherapy for ALCL, IMT, proliferativedisorders, glioblastoma and possible other solid tumors, or, as a singletherapeutic agent, could be used in a maintenance role to prevent cancerrecurrence in those patients. Various ALK inhibitors have been reported,such as indazoloisoquinolines (WO 2005/009389), thiazole amides andoxazole amides (WO 2005/097765), pyrrolopyrimidines (WO 2005080393), andpyrimidinediamines (WO 2005/016894).

In summary, there is clear genetic and biological evidence that linksaberrant ALK activation with the onset and progression of certain typesof cancer in humans. Considerable evidence indicates that ALK-positivetumor cells require these oncogenes to proliferate and survive, whileinhibition of ALK signaling leads to tumor cell growth arrest orapoptosis, resulting in objective cytoreductive effects. ALK isminimally expressed in most normal tissues in the healthy adult and areactivated and/or dysregulated in specific cancers during oncogenesisand/or during early stages of malignant progression. Consequently, theon-target effects of treatment with an ALK inhibitor against normalcells should be minimal, creating a favorable therapeutic index.

SUMMARY

This application provides compounds of the general formula (I)

and/or a salt thereof.

Compounds of formula (I) have ALK inhibitory activity, and may be usedto treat ALK mediated disorders or conditions.

This application further provides pharmaceutical compositions comprisingat least one compound of formula (I) or a salt thereof together with atleast one pharmaceutically acceptable excipient.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts pharmacokinetic/pharmacodynamic data from studies in SCIDmice bearing Sup-M2 human anaplastic large cell lymphoma (ALCL)xenografts dosed orally with select compounds described herein.

FIG. 2 depicts the anti-tumor efficacy of compound 16b (dosed orally,b.i.d.) in SCID mice bearing Sup-M2 ALCL xenografts.

FIG. 3 depicts the absolute stereochemical assignment of compound 17 viaheavy-atom anomalous dispersion x-ray crystallography.

DETAILED DESCRIPTION I. Definitions

As used herein, the following terms have the meanings ascribed to themunless specified otherwise.

The compounds and intermediates described herein may be named accordingto either the IUPAC (International Union for Pure and Applied Chemistry)or CAS (Chemical Abstracts Service) nomenclature systems.

The term “C_(x-y)” indicates the number of carbon atoms in a group. Forexample, a “C₁₋₆-alkyl” is an alkyl group having from one (1) to six (6)carbon atoms. In some instances, x=0, i.e., “C_(0-y)”. The term“C_(0-y)” indicates that the group may be absent or present, and ifpresent, defines the number of carbon atoms in the group. For example,“C₀₋₆-alkyl” indicates that an alkyl group may be absent (x=0) orpresent (x=1-6), and if present contains from one (1) to six (6) carbonatoms. For example, “—C₀₋₆-alkyl-C(═O)—C₀₋₆-alkyl-” includes —C(═O)—,—C₁₋₆-alkyl-C(═O)—, and —C₁₋₆-alkyl-C(═O)—C₁₋₆-alkyl-. Examples of—C₀₋₆-alkyl-C(═O)—C₀₋₆-alkyl- include, but are not limited to, —C(═O)—,—CH₂CH₂—C(═O)—, and —CH(CH₃)CH₂CH₂—C(═O)—CH₂—.

As used by themselves or in conjunction with another term or terms,“alkyl” or “alkyl group” refers to a monoradical of a branched orunbranched saturated hydrocarbon chain. Examples include, but are notlimited to, methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl,n-heptyl, n-octyl, n-nonyl, n-decyl, isopropyl, tert-butyl, isobutyl,etc. Alkyl groups typically contain 1-10 carbon atoms, such as 1-6carbon atoms, and can be substituted or unsubstituted.

As used by itself or in conjunction with another term or terms,“alkyloxy” refers to straight or branched hydrocarbon groups containingthe requisite number of carbon atoms (an alkyl group) as describedabove, bonded to an oxygen atom. As used herein, alkyloxy groups may beoptionally substituted with between one to four substituents.Representative examples of alkyloxy groups include, but are not limitedto, e.g. methoxy, ethoxy, tert-butoxy, etc.

“Halogen” includes fluorine, chlorine, bromine and iodine atoms.

“Pharmaceutical composition” refers to a composition having asafety/efficacy profile suitable for administration to a human.

“Pharmaceutically acceptable excipient” refers to physiologicallytolerable materials, which do not typically produce an allergic or otheruntoward reaction, such as gastric upset, dizziness and the like, whenadministered to a human.

“Pharmaceutically acceptable salt” refers to a salt having asafety/efficacy profile suitable for administration to a human.

“Subject” refers to a member of the class Mammalia. Examples of mammalsinclude, without limitation, humans, primates, chimpanzees, rodents,mice, rats, rabbits, horses, livestock, dogs, cats, sheep, and cows.

“Therapeutically effective amount” refers to an amount of a compoundsufficient to improve or inhibit worsening of symptoms associated with adisorder or condition being treated in a particular subject or subjectpopulation. It should be appreciated that determination of proper dosageforms, dosage amounts, and routes of administration is within the levelof ordinary skill in the pharmaceutical and medical arts. For example ina human or other mammal, a therapeutically effective amount can bedetermined experimentally in a laboratory or clinical setting, or may bethe amount required by the guidelines of the United States Food and DrugAdministration, or equivalent foreign agency, for the particular diseaseand subject being treated.

“Treatment” refers to the acute or prophylactic diminishment oralleviation of at least one symptom or characteristic associated orcaused by a disorder being treated. For example, treatment can includediminishment of a symptom of a disorder or complete eradication of adisorder.

“Administering” refers to the method of contacting a compound with asubject. Modes of “administering” include, but are not limited to,methods that involve contacting the compound intravenously,intraperitoneally, intranasally, transdermally, topically, viaimplantation, subcutaneously, parentally, intramuscularly, orally,systemically, and via adsorption.

For the purposes of this application, the term “and/or” should beunderstood as designating alternatives (this “or” that) as well ascombinations (this “and” that).

II. Compounds

This application provides compounds of the general formula (I)

and/or salt thereof, wherein:

X is CH or N;

R¹ is chosen from H or C₁-C₆alkyl substituted with at least one R³;at least one R² is halogen and the other R² is chosen from hydrogen orhalogen;each R³ is independently chosen from hydroxyl, C₁-C₆alkyoxy,—(CO)N(R⁴)₂, and —O(CO)R⁴; andeach R⁴ is independently chosen from hydrogen, C₁-C₆alkyl, and phenylsubstituted with at least one halogen.

In some embodiments, at least one R² is a halogen and the other R² ishydrogen. In other embodiments, both R² groups are halogen. In furtherembodiments, at least one R² is fluorine. In still further embodimentsboth R² groups are fluorine.

In additional embodiments X is CH. In still other embodiments X is N. Inyet further embodiments X is CH and at least one R² is a halogen and theother R² is hydrogen and/or X is CH and both R² groups are halogen. Instill other embodiments X is N and at least one R² is a halogen and theother R² is hydrogen and/or X is N and both R² groups are halogen.

In some embodiments, R¹ is H. In other embodiments, R¹ is H, X is N andat least one R² is a halogen and the other R² is hydrogen and/or R¹ isH, X is N and both R² groups are halogen. In some other embodiments, R¹is H, X is CH and at least one R² is a halogen and the other R² ishydrogen and/or R¹ is H, X is CH and both R² groups are halogen.

In other embodiments, R¹ is C₁-C₆alkyl optionally substituted with atleast one R³. In some embodiments, R³ is hydroxyl. In other embodiments,R³ is C₁-C₆alkyoxy. In yet other embodiments, R³ is —(CO)N(R⁴)₂. Instill further embodiments, R³ is —O(CO)R⁴.

In further embodiments, R⁴ is hydrogen. In yet further embodiments, R⁴is C₁-C₆alkyl. In additional embodiments, R⁴ is phenyl substituted withat least one halogen. In still other embodiments, R⁴ is phenylsubstituted with at least one bromine.

It should be understood that all stereochemical configurations,including individual enantiomers, diastereoisomers and/or mixtures ofstereoisomers (in any and all ratios), are included in the descriptionof formula (I). In particular, this application provides compounds offormula (I) that may be isolated as racemic diastereomers (namely the3,4-Cis or 3,4-Trans stereoisomers) as well as the single enantiomers ofsuch compounds, and any and all mixtures thereof. For example, thisapplication provides compounds of formula (I), or a salt thereof, whereR² is not hydrogen having any one of the following configurations:

In some embodiments, these compounds are isolated as single enantiomers.

In still other additional embodiments, this application providescompounds that are chosen from:

-   (±)-3,4-cis-1-(2-Methoxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol;-   (±)-3,4-cis-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol;-   (±)-2-(3,4-trans-3-Fluoro-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,    1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol;-   (±)-3,4-cis-4-{3-Methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol;-   (±)-[4-(3,4-trans-3-Fluoro-piperidin-4-yl)-2-methoxy-phenyl]-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-yl]-amine;-   (±)-2-(3,4-trans-3-fluoro-4-{3-methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol;-   (±)-2-(3,3-Difluoro-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol;-   (±)-1-(3,4-trans-3-Fluoro-4-{3-methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-2-methyl-propan-2-ol;-   (±)-2-(3,3-Difluoro-4-{3-methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol;-   (±)-[4-(3,3-Difluoro-piperidin-4-yl)-2-methoxy-phenyl]-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-yl]-amine;-   (±)-[4-(3,4-trans-3-Fluoro-piperidin-4-yl)-2-methoxy-phenyl]-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-yl]-amine;-   (3R,4S)-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol;-   (3S,4R)-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol;-   2-((3S,4S)-3-Fluoro-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol;-   2-((3R,4R)-3-Fluoro-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol;-   (3S,4S)-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,    1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol;-   (3R,4R)-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol;-   4-Bromo-benzoic acid    2-((3R,4R)-3-hydroxy-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethyl    ester;-   2-(4-{3-Methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol;    -   and/or a salt thereof.

In other embodiments, this application provides additional compoundsthat are isolated as single enantiomers, such as, for example,

and/or a salt thereof.

This application also provides salts of the compounds described herein.Preferably, the salts are pharmaceutically acceptable. Pharmaceuticallyacceptable acid addition salts of the compounds of formula I include,but are not limited to, salts derived from inorganic acids such ashydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydriodic, andphosphorus, as well as the salts derived from organic acids, such asaliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoicacids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, andaliphatic and aromatic sulfonic acids. Such salts thus include, but arenot limited to, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite,nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate,metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate,propionate, caprylate, isobutyrate, oxalate, malonate, succinate,suberate, sebacate, fumarate, maleate, mandelate, benzoate,chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate,benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate,maleate, tartrate, and methanesulfonate. Also contemplated are the saltsof amino acids such as arginate, gluconate, galacturonate, and the like;see, for example, Berge et al., “Pharmaceutical Salts,” J. ofPharmaceutical Science, 1977; 66:1-19.

The acid addition salts of the basic compounds may be prepared bycontacting the free base form with a sufficient amount of the desiredacid to produce the salt in the conventional manner. The free base formmay be regenerated by contacting the salt form with a base and isolatingthe free base in the conventional manner. The free base forms differfrom their respective salt forms somewhat in certain physical propertiessuch as solubility in polar solvents, but otherwise the salts are ingeneral equivalent to their respective free base for purposes of thepresent application.

Pharmaceutically acceptable base addition salts of compounds of formulaI are formed with metals or amines, such as alkali and alkaline earthmetal hydroxides, or of organic amines. Examples of metals used ascations include, but are not limited to, sodium, potassium, magnesium,and calcium. Examples of suitable amines include, but are not limitedto, N,N′-dibenzylethylenediamine, chloroprocaine, choline,diethanolamine, ethylenediamine (ethane-1,2-diamine), N-methylglucamine,and procaine; see, for example, Berge et al., supra., 1977.

The base addition salts of acidic compounds may be prepared bycontacting the free acid form with a sufficient amount of the desiredbase to produce the salt in the conventional manner. The free acid formmay be regenerated by contacting the salt form with an acid andisolating the free acid in a conventional manner. The free acid formsdiffer from their respective salt forms somewhat in certain physicalproperties such as solubility in polar solvents, but otherwise the saltsare in general equivalent to their respective free acid for purposes ofthe present application.

III. Pharmaceutical Compositions

The present application further provides pharmaceutical compositionscomprising a compound described herein (e.g., a compound of formula I ora pharmaceutically acceptable salt thereof, or any other compounddescribed in this application including compounds depicted in thespecification, examples or schemes), together with a pharmaceuticallyacceptable carrier, diluent, or excipient. The pharmaceuticalcomposition may contain two or more compounds (i.e., two or morecompounds may be used together in the pharmaceutical composition).Preferably, the pharmaceutical composition contains a therapeuticallyeffective amount of at least one compound of the present application. Inanother embodiment, these compositions are useful in the treatment of anALK-mediated disorder or condition. The compounds of the application canalso be combined in a pharmaceutical composition that also comprisescompounds that are useful for the treatment of cancer or anotherALK-mediated disorder.

A compound of the present application can be formulated as apharmaceutical composition in the form of a syrup, an elixir, asuspension, a powder, a granule, a tablet, a capsule, a lozenge, atroche, an aqueous solution, a cream, an ointment, a lotion, a gel, anemulsion, etc. Preferably, a compound of the present application willcause a decrease in symptoms or a disease indicia associated with an ALKmediated disorder as measured quantitatively or qualitatively.

For preparing a pharmaceutical composition using a compound of thepresent application, pharmaceutically acceptable carriers can be eithersolid or liquid. Solid form preparations include powders, tablets,pills, capsules, cachets, suppositories, and dispersible granules. Asolid carrier can be one or more substances which may also act asdiluents, flavoring agents, binders, preservatives, tabletdisintegrating agents, or an encapsulating material.

In powders, the carrier is a finely divided solid which is in a mixturewith the finely divided active component (i.e., compound of the presentapplication). In tablets, the active component is mixed with the carrierhaving the necessary binding properties in suitable proportions andcompacted in the shape and size desired.

The powders and tablets contain from 1% to 95% (w/w) of the activecompound (i.e., compound of the present application). In anotherembodiment, the active compound ranges from 5% to 70% (w/w). Suitablecarriers are magnesium carbonate, magnesium stearate, talc, sugar,lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose,sodium carboxymethylcellulose, a low melting wax, cocoa butter, and thelike. The term “preparation” is intended to include the formulation ofthe active compound with encapsulating material as a carrier providing acapsule in which the active component with or without other carriers, issurrounded by a carrier, which is thus in association with it.Similarly, cachets and lozenges are included. Tablets, powders,capsules, pills, cachets, and lozenges can be used as solid dosage formssuitable for oral administration.

For preparing suppositories, a low melting wax, such as a mixture offatty acid glycerides or cocoa butter, is first melted and the activecomponent is dispersed homogeneously therein, as by stirring. The moltenhomogeneous mixture is then poured into convenient sized molds, allowedto cool, and thereby to solidify.

Liquid form preparations include solutions, suspensions, and emulsions,for example, water or water/propylene glycol solutions. For parenteralinjection, liquid preparations can be formulated in solution in aqueouspolyethylene glycol solution.

Aqueous solutions suitable for oral use can be prepared by dissolvingthe active component in water and adding suitable colorants, flavors,stabilizers, and thickening agents as desired. Aqueous suspensionssuitable for oral use can be made by dispersing the finely dividedactive component in water with viscous material, such as natural orsynthetic gums, resins, methylcellulose, sodium carboxymethylcellulose,and other well-known suspending agents.

Also included are solid form preparations which are intended to beconverted, shortly before use, to liquid form preparations for oraladministration. Such liquid forms include solutions, suspensions, andemulsions. These preparations may contain, in addition to the activecomponent, colorants, flavors, stabilizers, buffers, artificial andnatural sweeteners, dispersants, thickeners, solubilizing agents, andthe like.

The pharmaceutical preparation is preferably in unit dosage form. Insuch form the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, such as packeted tablets, capsules, and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,or lozenge itself, or it can be the appropriate number of any of thesein packaged form.

The quantity of active component in a unit dose preparation may bevaried or adjusted from 0.1 mg to 1000 mg, preferably 1.0 mg to 100 mg,or from 1% to 95% (w/w) of a unit dose, according to the particularapplication and the potency of the active component. The compositioncan, if desired, also contain other compatible therapeutic agents.

Pharmaceutically acceptable carriers are determined in part by theparticular composition being administered, as well as by the particularmethod used to administer the composition. Accordingly, there is a widevariety of suitable formulations of pharmaceutical compositions of thepresent application (see, e.g., Remington: The Science and Practice ofPharmacy, 20th ed., Gennaro et al. Eds., Lippincott Williams andWilkins, 2000).

A compound of the present application, alone or in combination withother suitable components, can be made into aerosol formulations (i.e.,they can be “nebulized”) to be administered via inhalation. Aerosolformulations can be placed into pressurized acceptable propellants, suchas dichlorodifluoromethane, propane, nitrogen, and the like.

Formulations suitable for parenteral administration, such as, forexample, by intravenous, intramuscular, intradermal, and subcutaneousroutes, include aqueous and non-aqueous, isotonic sterile injectionsolutions, which can contain antioxidants, buffers, bacteriostats, andsolutes that render the formulation isotonic with the blood of theintended recipient, and aqueous and nonaqueous sterile suspensions thatcan include suspending agents, solubilizers, thickening agents,stabilizers, and preservatives. In the practice of this application,compositions can be administered, for example, by intravenous infusion,orally, topically, intraperitoneally, intravesically or intrathecally.The formulations of compounds can be presented in unit-dose ormulti-dose sealed containers, such as ampoules and vials. Injectionsolutions and suspensions can be prepared from sterile powders,granules, and tablets of the kind previously described.

The dose administered to a subject, in the context of the presentapplication should be sufficient to effect a beneficial therapeuticresponse in the subject over time. The dose will be determined by theefficacy of the particular compound employed and the condition of thesubject, as well as the body weight or surface area of the subject to betreated. The size of the dose also will be determined by the existence,nature, and extent of any adverse side-effects that accompany theadministration of a particular compound in a particular subject. Indetermining the effective amount of the compound to be administered inthe treatment or prophylaxis of the disorder being treated, thephysician can evaluate factors such as the circulating plasma levels ofthe compound, compound toxicities, and/or the progression of thedisease, etc. In general, the dose equivalent of a compound is fromabout 1 μg/kg to 10 mg/kg for a typical subject. Many differentadministration methods are known to those of skill in the art.

For administration, compounds of the present application can beadministered at a rate determined by factors that can include, but arenot limited to, the LD₅₀ of the compound, the pharmacokinetic profile ofthe compound, contraindicated drugs, and the side-effects of thecompound at various concentrations, as applied to the mass and overallhealth of the subject. Administration can be accomplished via single ordivided doses.

IV. Methods of Treatment

In another aspect, the present application provides a method of treatinga subject suffering from an ALK-mediated disorder or conditioncomprising: administering to the subject a therapeutically effectiveamount of a compound of formula I or a pharmaceutically acceptable saltform thereof. In another aspect, the present application provides acompound of formula I or a pharmaceutically acceptable salt form thereoffor use in treating a subject suffering from an ALK-mediated disorder orcondition. Preferably, the compound of formula I or a pharmaceuticallyacceptable salt form thereof is administered to the subject in apharmaceutical composition comprising a pharmaceutically acceptablecarrier. In another aspect, the present application provides apharmaceutical composition comprising a compound of formula I or apharmaceutically acceptable salt form thereof for use in treating asubject suffering from an ALK-mediated disorder or condition. In anotherembodiment, the ALK mediated condition or disorder is cancer. In anotherembodiment, the ALK-mediated condition is selected from anaplastic largecell lymphoma, inflammatory myofibroblastic tumor, glioblastoma, andother solid tumors. In another embodiment, the ALK-mediated condition isselected from colon cancer, breast cancer, renal cancer, lung cancer,hemangioma, squamous cell myeloid leukemia, melanoma, glioblastoma, andastrocytoma.

The ALK-mediated disorder or condition can be treated prophylactically,acutely, and chronically using compounds of the present application,depending on the nature of the disorder or condition. Typically, thehost or subject in each of these methods is human, although othermammals can also benefit from the administration of a compound of thepresent application.

In another embodiment, the present application provides a method oftreating a proliferative disorder in a subject, comprising administeringto the subject a therapeutically effective amount of a compound offormula I or a pharmaceutically acceptable salt form thereof. In anotheraspect, the present application provides a compound of formula I or apharmaceutically acceptable salt form thereof for use in treating aproliferative disorder in a subject. Preferably, the compound of formulaI or a pharmaceutically acceptable salt form thereof is administered tothe subject in a pharmaceutical composition comprising apharmaceutically acceptable carrier. In another aspect, the presentapplication provides a pharmaceutical composition comprising a compoundof formula I or a pharmaceutically acceptable salt form thereof for usein treating a proliferative disorder in a subject. In certainembodiments, the proliferative disorder is ALK mediated. In certainembodiments, the proliferative disorder is cancer. In certainembodiments, the proliferative disorder is selected from anaplasticlarge cell lymphoma, inflammatory myofibroblastic tumor, glioblastoma,and other solid tumors. In certain embodiments, the prolifereativedisorder is selected from colon cancer, breast cancer, renal cancer,lung cancer, hemangioma, squamous cell myeloid leukemia, melanoma,glioblastoma, and astrocytoma.

The proliferative disorder can be treated prophylactically, acutely, andchronically using compounds of the present application, depending on thenature of the disorder or condition. Typically, the host or subject ineach of these methods is human, although other mammals can also benefitfrom the administration of a compound of the present application.

The compounds of formula I share a common utility in treating ALKmediated disorders and a common core structure essential to that utility(i.e., the compounds of formula I are all pyrrolo[2,1-f][1,2,4]triazinederivatives).

In therapeutic applications, the compounds of the present applicationcan be prepared and administered in a wide variety of oral andparenteral dosage forms. Thus, the compounds of the present applicationcan be administered by injection, that is, intravenously,intramuscularly, intracutaneously, subcutaneously, intraduodenally, orintraperitoneally. Also, the compounds described herein can beadministered by inhalation, for example, intranasally. Additionally, thecompounds of the present application can be administered transdermally.In another embodiment, the compounds of the present application aredelivered orally. The compounds can also be delivered rectally, bucallyor by insufflation.

The compounds utilized in the pharmaceutical method of the applicationcan be administered at the initial dosage of about 0.001 mg/kg to about100 mg/kg daily. In another embodiment, the daily dose range is fromabout 0.1 mg/kg to about 10 mg/kg. The dosages, however, may be varieddepending upon the requirements of the subject, the severity of thecondition being treated, and the compound being employed. Determinationof the proper dosage for a particular situation is within the skill ofthe practitioner. Generally, treatment is initiated with smaller dosageswhich are less than the optimum dose of the compound. Thereafter, thedosage is increased by small increments until the optimum effect underthe circumstances is reached. For convenience, the total daily dosagemay be divided and administered in portions during the day, if desired.

V. Chemistry

Unless otherwise indicated, all reagents and solvents were obtained fromcommercial sources and used as received. ¹H NMRs were obtained on aBruker Avance at 400 MHz in the solvent indicated with tetramethylsilaneas an internal standard. Analytical HPLC was run using a Zorbax RX-C8,5×150 mm column eluting with a mixture of acetonitrile and watercontaining 0.1% trifluoroacetic acid with a gradient of 10-100%. LCMSresults were obtained on either of two instruments. First, in Examplesthat indicate LCMS retention times, analysis was performed on a WatersAquity Ultra Performance LC with a 2.1 mm×50 mm Waters Aquity UPLC BEHC18 1.7 m column. The target column temperature was 45° C., with a runtime of two (2) minutes, a flow rate of 0.600 mL/min, and a solventmixture of 5% (0.1% formic acid/water):95% (acetonitrile/0.1% formicacid). The mass spectrometry data was acquired on a Micromass LC-ZQ 2000quadrupole mass spectrometer. Second, in Examples that do not indicateLCMS retention times, analysis was performed on a Bruker Esquire 200 iontrap. Automated column chromatography was performed on a CombiFlashCompanion (ISCO, Inc.). Melting points were taken on a Mel-Tempapparatus and are uncorrected.

Synthesis

The compounds of the present application can be prepared in a number ofways well known to one skilled in the art of organic synthesis. Thecompounds of the present application can be synthesized using themethods described below as well as using alternate methods known to oneskilled in the art of organic chemistry including variations thereon asappreciated by those skilled in the art. The preferred methods include,but are not limited to or by, those described below. Unless otherwisestated, compounds are of commercial origin or readily synthesized bystandard methods well known to one skilled in the art of organicsynthesis.

The compounds of this application may be prepared using the reactionsand techniques described in this section. The reactions are performed insolvents appropriate to the reagents, and materials employed aresuitable for the transformations being effected. Also, in thedescription of the synthetic methods below, it is to be understood thatall proposed reaction conditions, including choice of solvent, reactionatmosphere, reaction temperature, duration of experiment and workupprocedures are chosen to be conditions standard for that reaction whichshould be readily recognized by one skilled in the art of organicsynthesis.

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application and thescope of the appended claims. Specific chemical transformations arelisted in the ensuing schemes and one skilled in the art appreciatesthat a variety of different reagents may be used in place of thoselisted. Common replacements for such reagents can be found in, but notlimited to, texts such as “Encyclopedia of Reagents for OrganicSynthesis” Leo A. Paquette, John Wiley & Son Ltd (1995) or“Comprehensive Organic Transformations: A Guide to Functional GroupPreparations” Richard C. Larock. Wiley-VCH and “Strategic Applicationsof Named Reactions in Organic Synthesis” Kurti and Czako, Elsevier, 2005and references therein.

General Reaction Schemes.

The synthesis of Examples 1, 2, 4, and 14 followed the general reactionsequence depicted in the general Scheme 1:

The synthesis of Examples 3, 5, 8, 10, and 13 followed the generalreaction sequence depicted in the general Scheme 2:

The synthesis of Examples 9, 11, and 12 followed the general reactionsequence depicted in the general Scheme 3:

The synthesis of Example 16 followed the general reaction sequencedepicted in Scheme 4:

The synthesis of Example 17 from Example 16b followed the reactionsequence outlined in Scheme 5:

EXPERIMENTAL PROCEDURES

Example 1(±)-3,4-cis-1-(2-Methoxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol

1a) 4-Chloro-2-methoxy-1-nitro-benzene (1.12 g, 5.96 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylicacid tert-butyl ester (1.84 g, 5.95 mmol),tetrakis(triphenylphosphine)palladium(0) (380 mg, 0.33 mmol), a 2 Msolution of potassium bicarbonate in water (7.45 mL, 14.9 mmol), and1,4-dioxane (18 mL) were combined in a sealed tube, and the mixture wasstirred and heated at 80° C. overnight. HPLC indicated the completeconversion of the starting material. The product,4-(3-methoxy-4-nitro-phenyl)-3,6-dihydro-2H-pyridine-1-carboxylic acidtert-butyl ester was isolated by flash chromatography (Silicagel,EtOAc/Hexanes 25-50%) in 90% yield.

1d) 4-(3-methoxy-4-nitro-phenyl)-3,6-dihydro-2H-pyridine-1-carboxylicacid tert-butyl ester, Intermediate 1a, was converted to(±)-(3R,4S)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol(Intermediate 1d) as outlined in Scheme 1 and described in detail inU.S. Pat. No. 8,471,005 (Appl. WO2010071885).

1e) A mixture of(±)-(3R,4S)-4-{3-Methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol(Intermediate 1d, prepared as described in U.S. Pat. No. 8,471,005/Appl.WO2010071885; 60 mg, 0.13 mmol), 1-bromo-2-methoxy-ethane, (26 mg, 0.19mmol), and sodium bicarbonate (13.6 mg, 0.16 mmol) in acetonitrile (1.5mL) was heated to reflux overnight, then the solvent was evaporatedunder vacuum and the title product was isolated by preparative reversephase hplc (on Gilson) as a yellow foam (14.5 mg, 21.4% yield). ¹H NMR(CDCl3): 8.68 (s, 1H), 8.30 (d, J=8.3 Hz, 1H), 7.99 (d, J=7.6 Hz, 1H),7.46 (br s, 1H), 7.42 (m, 1H), 7.12 (m, 1H), 7.06 (d, J=8.3 Hz, 1H),7.01 (d, J=4.1 Hz, 1H), 6.89 (s, 1H), 6.83 (d, J=4.1 Hz, 1H), 6.78 (d,J=8.2 Hz, 1H), 3.90 (br s, 1H), 3.89 (s, 3H), 3.84 (s, 3H), 3.54 (m,2H), 3.36 (s, 3H), 3.13 (m, 1H), 3.06 (m, 1H), 2.64 (m, 4H), 2.35 (d,J=11.4 Hz, 1H), 2.22 (br s, 2H), 1.68 (m, 1H); LC/MS (ESI+): 504.1(M+H).

Example 2(±)-3,4-cis-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol

Ethylene oxide (40 mg, 0.9 mmol) was trapped in a glass tube at −78° C.and then(±)-(3R,4S)-4-{3-Methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol(Example 4; 40 mg, 0.09 mmol) in tetrahydrofuran (200 L) was added atthe same temperature and the tube was capped and the reaction wasallowed to stir overnight at room temperature. The volatiles were thenevaporated under reduced pressure and the product was isolated by (Isco)flash column chromatography (DCM/MeOH on Silicagel) to afford theproduct as a yellow foam (25 mg, 57% yield). ¹H NMR (CDCl₃): 8.70 (s,1H), 8.48 (d, J=7.6 Hz, 1H), 8.24 (d, J=8.1 Hz, 1H), 7.47 (br s, 1H),7.13 (d, J=4.1 Hz, 1H), 7.07 (m, 2H), 6.88 (s, 1H), 6.82 (m, 2H), 4.02(s, 3H), 3.96 (br s, 1H), 3.90 (s, 3H), 3.70 (m, 2H), 3.11 (m, 2H), 2.64(m, 3H), 2.38 (d, J=11.5 Hz, 1H), 2.22 (m, 4H), 1.72 (d, J=11.5 Hz, 1H);LC/MS (ESI+): 491.0 (M+H)

Example 3(±)-2-(3,4-trans-3-Fluoro-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol

(±)-[4-(3,4-Trans-3-fluoro-piperidin-4-yl)-2-methoxy-phenyl]-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-yl]-amine(Example 5; 55 mg, 0.12 mmol) was converted to the title product by aprocedure similar to the one described for Example 2 to give a yellowfoam (35 mg, 58% yield). ¹H NMR (CDCl₃): 8.69 (s, 1H), 8.34 (d, J=8.0Hz, 1H), 7.98 (d, J=7.6 Hz, 1H), 7.48 (s, 1H), 7.44 (m, 1H), 7.14 (m,1H), 7.07 (d, J=8.3 Hz, 1H), 7.02 (d, J=4.1 Hz, 1H), 6.84 (d, J=4.1 Hz,1H), 6.77 (m, 2H), 4.63 (dddd, J=48.4; 9.7; 9.7; 4.6 Hz, 1H), 3.90 (s,3H), 3.84 (s, 3H), 3.67 (m, 2H), 3.35 (m, 1H), 2.96 (d, J=11.4 Hz, 1H),2.65 (m, 3H), 2.55 (br s, 1H; —OH), 2.19 (m, 2H), 1.86 (m, 2H); LC/MS(ESI+): 492.0 (M+H).

Example 4(±)-3,4-cis-4-{3-Methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol

Into a 8-dram vial,7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ol (preparedas described in U.S. Pat. No. 8,471,005/Appl. WO2010071885; 70 mg, 0.29mmol), N,N-dimethylformamide (1. mL, 13 mmol), N,N-diisopropylethylamine(0.15 mL) and N-phenylbis(trifluoromethanesulphonimide) (124 mg, 0.35mmol) were added. The reaction mixture was stirred at room temperaturefor one hour.(±)-(3R,4S)-4-(4-Amino-3-methoxy-phenyl)-3-hydroxy-piperidine-1-carboxylicacid tert-butyl ester (Intermediate 12P prepared as described in Scheme2 and in U.S. Pat. No. 8,471,005/Appl. WO2010071885; 111.8 mg, 0.35mmol) was added. The reaction was heated at 80° C. overnight. Thereaction mixture was partitioned between EtOAc and aqueous sodiumbicarbonate, the organic extracts were dried (MgSO₄) and the solvent wasevaporated under reduced pressure. The crude residue was taken inmethylene chloride (1 mL) and treated with trifluoroacetic acid (0.45mL, 5.78 mmol) at room temperature until HPLC indicated complete removalof the Boc group. The reaction mixture was concentrated under vacuum andthe product was isolated by reverse-phase pre-hplc (Gilson) followed byfree base isolation by work-up on catch/release acid-resin(Strata/Phenomenex) to provide the product as a yellow foam (82.00 mg,63.5%). ¹H NMR (CDCl₃): 8.72 (s, 1H), 8.48 (d, J=7.6 Hz, 1H), 8.25 (m,2H), 7.48 (s, 1H), 7.14 (d, J=4.1 Hz, 1H), 7.08 (m, 1H), 6.83 (m, 4H),4.02 (s, 3H), 3.92 (s, 3H), 3.88 (br s, 1H), 3.24 (m, 2H), 2.89 (d,J=12.6 Hz, 1H), 2.76 (m, 2H), 2.17 (m, 2H), 1.64 (d, J=12.6 Hz, 1H);LC/MS (ESI+): 447.0 (M+H).

Example 5(i)-[4-(3,4-trans-3-Fluoro-piperidin-4-yl)-2-methoxy-phenyl]-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-yl]-amine

5a). A solution of(±)-3,4-trans-3-hydroxy-4-(3-methoxy-4-nitro-phenyl)-piperidine-1-carboxylicacid tert-butyl ester (Intermediate 16a, prepared as described in Scheme4 and in U.S. Pat. No. 8,471,005/Appl. WO2010071885; 200 mg, 0.57 mmol)in 1,2-dichloroethane (6 mL) was treated with diethylaminosulfurtrifluoride (188 uL, 1.42 mmol) at ˜0° C., and the reaction mixture wasstirred overnight, allowing it to warm to room temperature. The reactionmixture was quenched with dilute aq. NaHCO₃, and the product extractedwith DCM. The organic extracts were washed once with water, dried(MgSO₄), then filtered, and the solvent was evaporated under reducedpressure to afford the crude product,(±)-3-fluoro-4-(3-methoxy-4-nitro-phenyl)-piperidine-1-carboxylic acidtert-butyl ester (200 mg, 99% yield), which was used withoutpurification. ¹H NMR (CDCl₃): δ ppm 7.86 (d, J=8.1 Hz, 1H), 6.87-7.00(m, 2H), 4.58 (br. s., 2H), 4.13-4.32 (m, 1H), 3.98 (s, 3H), 2.70-2.92(m, 3H), 1.93 (d, J=13.6 Hz, 1H), 1.68-1.83 (m, 1H), 1.49 (s, 9H).

5b). To a solution of(±)-3-fluoro-4-(3-methoxy-4-nitro-phenyl)-piperidine-1-carboxylic acidtert-butyl ester in methanol (10 mL) was added palladium on carbon 10%(1:9 palladium:carbon black, 40 mg). The mixture was shaken in a Parrapparatus under an atmosphere of hydrogen (35 PSI) overnight. Filtrationthrough Celite and evaporation of the solvent provided crude(±)-4-(4-amino-3-methoxy-phenyl)-3-fluoro-piperidine-1-carboxylic acidtert-butyl ester (180 mg, 98%), which was used without purification. ¹HNMR (CDCl₃): δ ppm 6.63-6.71 (m, 3H), 4.52 (br. s., 2H), 4.05-4.26 (m,1H), 3.85 (s, 3H), 3.74 (br. s., 2H), 2.63-2.86 (m, 3H), 1.88 (d, J=13.4Hz, 1H), 1.62-1.79 (m, 1H), 1.49 (s, 9H).

5c). 7-(2-Methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ol (prepared asdescribed in U.S. Pat. No. 8,471,005/Appl. WO2010071885; 120 mg, 0.50mmol) and(±)-3,4-trans-4-(4-amino-3-methoxy-phenyl)-3-fluoro-piperidine-1-carboxylicacid tert-butyl ester (194 mg, 0.6 mmol) were converted to the titleproduct by a procedure similar to the one described for Example 4:yellow foam (111.00 mg, 50% yield). ¹H NMR (CDCl₃): 8.69 (s, 1H), 8.35(d, J=8.2 Hz, 1H), 7.98 (d, J=7.6 Hz, 1H), 7.47 (s, 1H), 7.43 (m, 1H),7.13 (m, 1H), 7.07 (d, J=8.3 Hz, 1H), 7.02 (d, J=4.3 Hz, 1H), 6.84 (d,J=4.3 Hz, 1H), 6.82 (m, 2H), 4.56 (dddd, J=48.5; 9.7; 9.7; 5.0 Hz, 1H),3.90 (s, 3H), 3.84 (s, 3H), 3.48 (d, J=11.3 Hz, 1H), 3.08 (d, J=11.8 Hz,1H), 2.68 (m, 3H), 1.92 (m, 1H), 1.76 (m, 2H); LC/MS (ESI+):448.0 (M+H).

Examples 6 and 72-((3S,4S)-3,4-Dihydroxy-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-acetamideand2-((3R,4R)-3,4-Dihydroxy-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-acetamide

Enantiomerically pure Examples 6 and 7 were prepared by chromatographicseparation from racemic(±)-2-(3,4-cis-3,4-dihydroxy-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-acetamide(exemplified in U.S. Pat. No. 8,471,005/Appl. WO2010071885) by a SFCprocedure similar to the one described for Examples 16a and 16b. Example6 (ee>99%) is first to elute from the column, and Example 7 is thesecond to elute from the column (ee>99%).

Example 8(±)-2-(3,4-trans-3-fluoro-4-{3-methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol

Ethylene oxide and(±)-[4-(3,4-trans-3-Fluoro-piperidin-4-yl)-2-methoxy-phenyl]-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-yl]-amine(Example 13, 60 mg) in methanol (0.4 mL) were converted to the titleproduct by a procedure similar to the one described for Example 2, butat 50° C., to give a yellow foam (34 mg, 52% yield). ¹H NMR (CDCl₃):8.72 (s, 1H), 8.48 (d, J=7.5 Hz, 1H), 8.29 (d, J=8.2 Hz, 1H), 8.25 (m,1H), 7.49 (br s, 1H), 7.14 (d, J=3.2 Hz, 1H), 7.08 (m, 1H), 6.82 (m,3H), 4.65 (dddd, J=48 (H—F coupling), 11.2, 11.2, 3.4 Hz, 1H), 4.02 (s,3H), 3.92 (s, 3H), 3.68 (m, 2H), 3.36 (m, 1H), 2.98 (d, J=11.0 Hz, 1H),2.67 (m, 3H), 2.22 (m, 3H), 1.92 (m, 1H), 1.84 (m, 1H); LC/MS (ESI+):493.0 (M+H).

Example 9(±)-2-(3,3-Difluoro-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol

(±)-[4-(3,3-Difluoro-piperidin-4-yl)-2-methoxy-phenyl]-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-yl]-amine(prepared in a manned similar to Example 12; 68 mg, 0.15 mmol) wasconverted to the title product by a procedure similar to the onedescribed for Example 8 to give a yellow foam (47 mg, 63% yield). ¹H NMR(CDCl₃): 8.70 (s, 1H), 8.34 (d, J=8.2 Hz, 1H), 7.97 (d, J=7.6 Hz, 1H),7.50 (s, 1H), 7.43 (m, 1H), 7.13 (m, 1H), 7.07 (d, J=8.3 Hz, 1H), 7.02(d, J=4.2 Hz, 1H), 6.82 (m, 3H), 3.91 (s, 3H), 3.84 (s, 3H), 3.68 (m,2H), 3.24 (m, 1H), 3.09 (m, 1H), 2.87 (m, 1H), 2.68 (m, 2H), 2.45 (dd,J=28.2 (F—H vicinal coupling), 11.6 Hz, 1H), 2.27 (m, 3H), 1.89 (m, 1H);LC/MS (ESI+): 509.9 (M+H).

Example 10(±)-1-(3,4-trans-3-Fluoro-4-{3-methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-2-methyl-propan-2-ol

1,2-Epoxy-2-methylpropane and(±)-[4-(3,4-trans-3-fluoro-piperidin-4-yl)-2-methoxy-phenyl]-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-yl]-amine(Example 13, 58 mg) were converted to the title product by a proceduresimilar to the one described for Example 8 to give a yellow foam (36 mg,53% yield). ¹H NMR (CDCl₃): 8.71 (s, 1H), 8.48 (d, J=7.4 Hz, 1H), 8.28(d, J=8.2 Hz, 1H), 8.26 (d, J=4.5 Hz, 1H), 7.49 (s, 1H), 7.13 (d, J=4.5Hz, 1H), 7.10 (m, 1H), 6.84 (d, J=4.5 Hz, 1H), 6.79 (m, 2H), 4.64 (dddd,J=48.3 (F—H gem. coupling), 9.8, 9.8, 4.3 Hz, 1H), 4.02 (s, 3H), 3.93(s, 3H), 3.36 (m, 1H), 2.96 (m, 1H), 2.86 (br s, 1H), 2.60 (m, 1H), 2.47(m, 4H), 1.87 (m, 2H), 1.22 (s, 3H), 1.21 (s, 3H); LC/MS (ESI+): 521.0(M+H).

Example 11(±)-2-(3,3-Difluoro-4-{3-methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol

Ethylene oxide and(±)-[4-(3,3-difluoro-piperidin-4-yl)-2-methoxy-phenyl]-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-yl]-amine(Example 12, 68 mg) were converted to the title product by a proceduresimilar to the one described for Example 8 to give a yellow foam (51 mg,68% yield). ¹H NMR (CDCl₃): 8.72 (s, 1H), 8.48 (d, J=7.4 Hz, 1H), 8.28(d, J=8.1 Hz, 1H), 8.24 (d, J=4.3 Hz, 1H), 7.51 (s, 1H), 7.14 (d, J=3.6Hz, 1H), 7.08 (m, 1H), 6.84 (m, 2H), 4.02 (s, 3H), 3.92 (s, 3H), 3.68(m, 2H), 3.25 (m, 1H), 3.10 (m, 1H), 2.90 (m, 1H), 2.74 (m, 1H), 2.65(m, 1H), 2.46 (dd, J=28.3 (F—H vic. coupling), 11.7 Hz, 1H), 2.27 (m,3H), 1.90 (m, 1H); LC/MS (ESI+): 510.54 (M+H).

Example 12(±)-[4-(3,3-Difluoro-piperidin-4-yl)-2-methoxy-phenyl]-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-yl]-amine

12a). 1-Bromo-3-methoxy-benzene (10 g, 53.5 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylicacid tert-butyl ester (16.5 g, 53.5 mmol),tetrakis(triphenylphosphine)palladium(0) (3.1 g, 2.7 mmol), a solutionof 2 M sodium carbonate in water (67 mL, 133.7 mmol), were combined in1,4-dioxane (150 mL) in a round bottom flask, and the mixture was heatedat 80° C. overnight. The product was isolated by flash chromatography(Silicagel, EtOAc/Hexanes) to afford4-(3-Methoxy-phenyl)-3,6-dihydro-2H-pyridine-1-carboxylic acidtert-butyl ester (12.5 g, 81% yield). ¹H NMR (CDCl₃): 7.26 (m, 1H), 6.97(d, J=7.6 Hz, 1H), 6.91 (s, 1H), 6.81 (d, J=8.1 Hz, 1H), 6.04 (br. s.,1H), 4.07 (br. s., 2H), 3.82 (s, 3H), 3.60-3.68 (m, 2H), 2.52 (br. s.,2H), 1.49 (s, 9H).

12b). 4-(3-Methoxy-phenyl)-3,6-dihydro-2H-pyridine-1-carboxylic acidtert-butyl ester (2 g, 6.9 mmol) was further purified by flashchromatography (100 DCM on Silicagel) to remove baseline impurities(likely waste palladium from Suzuki reaction), then was dissolved inmethylene chloride (28.9 mL, 451 mmol) and treated with m-CPBA 70-75%(70:30, m-chloroperbenzoic acid:3-chlorobenzoic acid, 2.4 g, 9.7 mmol)at room temperature overnight. The reaction was quenched by treatmentwith a Na₂S₂O₃ aq solution, followed by sat. NaHCO₃ aq. solution, andfurther extracted twice with dichloromethane. The combined organicextracts were dried (MgSO₄) and filtered, and the solvent was evaporatedunder reduce pressure. The product was used without further purificationin the next step:(±)-6-(3-methoxy-phenyl)-7-oxa-3-aza-bicyclo[4.1.0]heptane-3-carboxylicacid tert-butyl ester (2 g, 95% yield). ¹H NMR (CDCl₃): δ ppm 7.26 (m,1H), 6.95 (d, J=7.8 Hz, 1H), 6.90 (br. s., 1H), 6.84 (d, J=8.1 Hz, 1H),3.93-4.19 (m, 1H), 3.81 (s, 3H), 3.53-3.79 (m, 2H), 3.10-3.24 (m, 2H),2.38-2.51 (m, 1H), 2.17 (br. s., 1H), 1.48 (s, 9H).

12c). To a solution of(±)-6-(3-methoxy-phenyl)-7-oxa-3-aza-bicyclo[4.1.0]heptane-3-carboxylicacid tert-butyl ester (2 g) in methanol (115 mL) was added palladium oncarbon 10% (10:90, palladium:carbon black, 400 mg). The mixture wasshaken in a Parr apparatus under an atmosphere of hydrogen (50 PSI)overnight. Filtration through Celite and evaporation of the solventprovided the crude product. The product was purified by flashchromatography (Silicagel, EtOAc/hexanes) to give(±)-3-hydroxy-4-(3-methoxy-phenyl)-piperidine-1-carboxylic acidtert-butyl ester (1.79 g, 89% yield). ¹H NMR (CDCl₃): δ ppm 7.26 (s,1H), 6.72-6.92 (m, 3H), 4.31 (br. s., 2H), 3.96 (br. s., 1H), 3.81 (s,3H), 3.01 (d, J=13.4 Hz, 1H), 2.70-2.92 (m, 2H), 2.13-2.34 (m, 1H), 1.64(br. s., 2H), 1.49 (s, 9H).

12d). (±)-3-Hydroxy-4-(3-methoxy-phenyl)-piperidine-1-carboxylic acidtert-butyl ester (1.27 g, 4.13 mmol) and pyridine (334 uL, 4 mmol) inmethylene chloride (70 mL) at 0° C. were treated with Dess-Martinperiodinane (3.5 g, 8.3 mmol) and then the reaction was stirred at roomtemperature for 2 h. The reaction was quenched by addition of a mixtureof sat. solution of Na₂S₂O₃ and sat. solution of NaHCO₃ (1:1 v:v) (50ml), which was followed by extraction with ether. The product wasisolated by flash chromatography to afford(±)-4-(3-methoxy-phenyl)-3-oxo-piperidine-1-carboxylic acid tert-butylester (0.94 g, 75% yield). ¹H NMR (CDCl₃): δ ppm 7.27 (s, 1H), 6.83 (d,J=8.1 Hz, 1H), 6.70-6.75 (m, 1H), 6.68 (s, 1H), 4.24 (d, J=18.2 Hz, 1H),4.06 (d, J=18.2 Hz, 2H), 3.80 (s, 3H), 3.62 (dd, J=11.5, 5.9 Hz, 1H),3.51 (br. s., 1H), 2.19-2.35 (m, 2H), 1.44-1.53 (m, 9H).

12e). A solution of(±)-4-(3-methoxy-phenyl)-3-oxo-piperidine-1-carboxylic acid tert-butylester (900 mg, 2.95 mmol) in 1,2-dichloroethane (50 mL) was treated withdiethylaminosulfur trifluoride (974 uL, 7.4 mmol) at ˜0° C., and thereaction mixture was stirred overnight, allowing it to warm to roomtemperature. The reaction mixture was quenched with dilute aq. NaHCO₃,and the product was extracted with DCM. The organic extracts were washedonce with water, dried (MgSO₄) and filtered, then the solvent wasevaporated under reduced pressure. The crude product was used withoutpurification:(±)-3,3-difluoro-4-(3-methoxy-phenyl)-piperidine-1-carboxylic acidtert-butylester (515 mg, 53% yield). ¹H NMR (CDCl₃): δ ppm 7.22-7.30 (m,1H), 6.89 (d, J=7.6 Hz, 1H), 6.86 (br. s., 2H), 4.13-4.67 (m, 2H), 3.81(s, 3H), 2.92-3.17 (m, 2H), 2.83 (br. s., 1H), 2.09-2.24 (m, 1H), 1.87(d, J=12.4 Hz, 1H), 1.49 (s, 9H).

12f). Nitric acid (77 uL, 1.8 mmol) was added to acetic anhydride (865uL, 9.2 mmol) at 0° C. under an atmosphere of nitrogen and the mixturewas stirred for 5-10 min. The resulting solution was added to a solutionof (±)-3,3-difluoro-4-(3-methoxy-phenyl)-piperidine-1-carboxylic acidtert-butyl ester (500 mg, 1.5 mmol) in acetonitrile (12 mL) at 0° C.under an atmosphere of nitrogen, and the reaction was stirred for 2hours at this temperature. Cold DCM was added, then the reaction wasquenched with sat. aq. NaHCO₃. The organic extracts were dried (MgSO₄),filtered, and concentrated under reduced pressure. The desiredregioisomer product was isolated by flash chromatography (ISCO,Silicagel, EtOAc/Hexanes 1:4) to afford the(±)-3,3-difluoro-4-(3-methoxy-4-nitro-phenyl)-piperidine-1-carboxylicacid tert-butyl ester (240 mg, 42% yield) as a white foamy solid. ¹H NMRand NOEDiff (CDCl₃): δ ppm 7.84 (d, J=8.3 Hz, 1H), 7.02 (s, 1H), 6.97(d, J=8.3 Hz, 1H), 4.42 (br. s., 2H), 3.97 (s, 3H), 2.73-3.20 (m, 3H),2.09-2.27 (m, 1H), 1.91 (d, J=14.1 Hz, 1H), 1.50 (s, 9H).

12g). To a solution of(±)-3,3-difluoro-4-(3-methoxy-4-nitro-phenyl)-piperidine-1-carboxylicacid tert-butyl ester (230.00 mg) in methanol (10 mL) was addedpalladium on carbon 10% (10:90, palladium: carbon black, 40 mg). Themixture was shaken in a Parr apparatus under an atmosphere of hydrogen(35 PSI) for 2 h. Filtration through Celite and evaporation of thesolvent provided(±)-4-(4-amino-3-methoxy-phenyl)-3,3-difluoro-piperidine-1-carboxylicacid tert-butyl ester (200 mg, 95% yield), which was used withoutpurification. ¹H NMR (CDCl₃): δ ppm 6.65-6.77 (m, 3H), 4.15-4.65 (m,2H), 3.85 (s, 3H), 3.77 (br. s., 2H), 2.72-3.13 (m, 3H), 2.03-2.24 (m,1H), 1.85 (d, J=12.4 Hz, 1H), 1.49 (s, 9H) 3124-67

12h). 7-(2-Methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ol(prepared as described in U.S. Pat. No. 8,471,005/Appl. WO2010071885; 65mg, 0.27 mmol) and(±)-4-(4-amino-3-methoxy-phenyl)-3,3-difluoro-piperidine-1-carboxylicacid tert-butyl ester (100 mg, 0.29 mmol) were converted to the titlecompound by a procedure similar to Example 4, to afford a yellow foam(86 mg, 69% yield). ¹H NMR (CDCl₃): 8.72 (s, 1H), 8.49 (d, J=7.4 Hz,1H), 8.28 (d, J=8.2 Hz, 1H), 8.24 (m, 1H), 7.51 (s, 1H), 7.14 (d, J=4.0Hz, 1H), 7.08 (m, 1H), 6.86 (m, 3H), 4.02 (s, 3H), 3.92 (s, 3H), 3.30(m, 1H), 3.21 (m, 1H), 3.02 (m, 1H), 2.90 (dd, J=30.8 (F_H vic.coupling), 13.7 Hz, 1H), 2.75 (m, 1H), 2.10 (m, 1H), 1.95 (m, 2H); LC/MS(ESI+): 467.0 (M+H).

Example 13(±)-[4-(3,4-trans-3-Fluoro-piperidin-4-yl)-2-methoxy-phenyl]-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-yl]-amine

7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ol (preparedas described in U.S. Pat. No. 8,471,005/Appl. WO2010071885; 100 mg, 0.41mmol) and(±)-4-(4-amino-3-methoxy-phenyl)-3-fluoro-piperidine-1-carboxylic acidtert-butyl ester (Intermediate 5b, Scheme 2; 146 mg, 0.45 mmol) wereconverted to the title compound by a procedure similar to Example 4, toafford a yellow foam (119 mg, 64% yield). ¹H NMR (CDCl₃): δ ppm 8.70 (s,1H), 8.49 (d, J=7.6 Hz, 1H), 8.28 (d, J=8.1 Hz, 1H), 8.19-8.26 (m, 1H),7.48 (s, 1H), 7.13 (d, J=4.3 Hz, 1H), 7.01-7.09 (m, 1H), 6.77-6.87 (m,3H), 4.46-4.69 (m, 1H), 4.02 (s, 3H), 3.91 (s, 3H), 3.44-3.50 (m, 2H),3.08 (d, J=11.9 Hz, 1H), 2.60-2.80 (m, 3H), 1.68-1.83 (m, 2H).

Example 14(±)-3,4-cis-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol

Racemic Example 14,(±)-3,4-cis-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol,was prepared as described in U.S. Pat. No. 8,471,005/Appl. WO2010071885and in Scheme 1.

Examples 14a and 14b(3R,4S)-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-oland(3S,4R)-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol

Enantiomerically pure Examples 14a and 14b were prepared bychromatographic separation from the racemic Example 14 (exemplified inU.S. Pat. No. 8,471,005/Appl. WO2010071885) by a SFC procedure similarto the one described for Examples 16a and 16b. Example 14a (ee>99%) isfirst to elute from the column, and Example 14b is the second to elutefrom the column (ee>99%).

Examples 15a and 15b2-((3S,4S)-3-Fluoro-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanoland2-((3R,4R)-3-Fluoro-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol

Enantiomerically pure Examples 15a and 15b were prepared bychromatographic separation from the racemic Example 3 by a SFC proceduresimilar to the one described for Examples 16a and 16b. Example 15a(ee>99%) is first to elute from the column, and Example 15b is thesecond to elute from the column (ee>99%).

Example 16(±)-3,4-trans-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol

16a) The reaction was placed under an atmosphere of nitrogen. To a cold0° C. solution of4-(3-methoxy-4-nitro-phenyl)-3,6-dihydro-2H-pyridine-1-carboxylic acidtert-butyl ester (intermediate 4, 4.0 g, 12.0 mmol) in drydimethoxyethane (10 ml) was added sodium borohydride (728 mg, 19.2 mmol)in one portion. After 5 minutes a solution of boron trifluoride etherate(3.0 mL, 24 mmol) in dry dimethoxyethane (5 mL) was added dropwise(syringe), and then the cooling bath removed and the reaction wasstirred overnight (while warming to room temperature). The reaction wascooled to 0° C. and water was added dropwise until effervescence ceased(quench of excess hydride species), followed by a 10 M solution ofsodium hydroxide in water (7.6 mL, 76 mmol) and then 30% aq. hydrogenperoxide (7.6 mL, 74 mmol) dropwise. After the addition, the reactionwas allowed to warm to room temperature and stirred for 2 h. Dilutedwith water and then extracted with EtOAc. The combined extracts werewashed with 3% aq. NH₃, then water, dried (MgSO4), filtered, evaporated,and the residue purified by flash chromatography (ISCO, Silicagel,EtOAc/Hexanes 25-40%) to give 2 g (47% yield) of intermediate 16a,(±)-3,4-trans-3-hydroxy-4-(3-methoxy-4-nitro-phenyl)-piperidine-1-carboxylicacid tert-butylester.

16b). To a solution of(±)-3,4-trans-3-hydroxy-4-(3-methoxy-4-nitro-phenyl)-piperidine-1-carboxylicacid tert-butylester (intermediate 16a, 7 g) in methanol (350 mL) wasadded palladium on carbon 10% (10:90, Palladium:carbon black, 1 g). Themixture was shaken in a Parr apparatus under an atmosphere of hydrogen(35 PSI) overnight. Filtration through Celite and evaporation of thesolvent provided the crude product. The crude product was worked-up bysuspension in dichlomethane and washing twice with diluted aq. NaHCO₃,drying on Na₂SO₄, filtration and evaporation of solvent. The product waspurified by flash chromatography (Silicagel, EtOAc/hexanes ˜40-70%) togive 5.1 g (80% yield) of intermediate 16b,(±)-3,4-trans-4-(4-amino-3-methoxy-phenyl)-3-hydroxy-piperidine-1-carboxylicacidtert-butyl ester.

Intermediate 16b was converted to racemic Example 16 as outlined inScheme 4 and described in detail in U.S. Pat. No. 8,471,005 (Appl.WO2010071885).

Example 16a(3S,4S)-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-oland Example 16b(3R,4R)-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol

Separation of Enantiomers:

Examples 16a and 16b were obtained from Example 16 as follows: RacemicExample 16 (exemplified in U.S. Pat. No. 8,471,005/Appl. WO2010071885)was subjected to enantiomeric separation by supercritical fluid (SFC)high performance liquid chromatography on chiral stationary phase columnto generate isolated single enantiomer Examples 16a and 16b. A ChiralPakAD-H (10×150 mm or 21×150 mm) was used, at a temperature T=35° C. andback pressure P=120 bar, with a UV detector set at 220 nm wavelength.The flow rate was 6.0 mL/min and the mobile phase was 40% MeOH (with0.1% DEA)—60% CO₂. Example 16a (ee>99%) is first to elute from thecolumn, and Example 16b is the second to elute from the column (ee>99%).

Example 17 4-Bromo-benzoic acid2-((3R,4R)-3-hydroxy-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethylester

To a mixture of(3R,4R)-1-(2-hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol(Example 16b; 40.00 mg, 0.082 mmol) and triethylamine (12 uL, 0.09 mmol)in methylene chloride (0.400 mL, 6.24 mmol) was added 4-bromo-benzoylchloride, (19 mg, 0.086 mmol) at 0° C. The reaction was stirredovernight allowing the cooling bath/reaction to slowly warm to roomtemperature. The volatile were evaporated and the crude residue wasfurther dried on high vacuum, The product was isolated by preparativereverse phase hplc (Gilson) and the free base was released by using astrong cation exchange resin column (Strata, from Phenomenex) to give 33mg (60% yield) of 4-bromo-benzoic acid2-((3R,4R)-3-hydroxy-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethyl ester. ¹HNMR (CDCl₃): 8.70 (br s, 1H), 8.35 (d, J=8.1 Hz, 1H), 7.96 (d, J=7.6 Hz,1H), 7.92 (d, J=8.4 Hz, 2H), 7.60 (d, J=8.4 Hz, 2H), 7.47 (br s, 1H),7.46-7.40 (m, 1H), 7.12 (app t, J=7.5 Hz, 1H), 7.07 (d, J=8.3 Hz, 1H),7.02 (d, J=4.7 Hz, 1H), 6.85 (d, J=4.7 Hz, 1H), 6.78 (d, J=8.7 Hz, 1H),6.76 (br s, 1H), 4.50 (app t, J=5.8 Hz, 2H), 3.89 (s, 3H), 3.84 (s, 3H),3.80 (m, 1H), 3.31 (m, 1H), 3.04 (br d, J=11.3 Hz, 1H), 2.90 (app t,J=5.8 Hz, 2H), 2.35 (m, 1H), 2.26 (m, 1H), 2.15 (app t, J=10.2 Hz, 1H),1.86 (m, 2H), 1.67 (br s, 1H).

Example 182-(4-{3-Methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol

[7-(2-Methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-yl]-(2-methoxy-4-piperidin-4-yl-phenyl)-amine(60 mg, 0.14 mmol) was prepared in 60% yield from7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ol (described inU.S. Pat. No. 8,471,005/Appl. WO2010071885) and4-(4-amino-3-methoxy-phenyl)-piperidine-1-carboxylic acid tert-butylester in manner similar to Example 4, and converted to the titlecompound by a procedure similar to the one described for Example 8 togive a yellow foam (35 mg, 53% yield). ¹H NMR (CDCl₃): 8.69 (s, 1H),8.29 (d, J=7.7 Hz, 1H), 8.99 (d, J=7.6 Hz, 1H), 7.43 (m, 2H), 7.13 (m,1H), 7.07 (d, J=8.3 Hz, 1H), 7.02 (d, J=2.7 Hz, 1H), 6.83 (d, J=2.7 Hz,1H), 6.73 (s, 1H), 6.72 (m, 1H), 3.90 (s, 3H), 3.84 (s, 3H), 3.65 (m,2H), 3.05 (m, 2H), 2.59 (m, 2H), 2.48 (m, 1H), 2.19 (m, 2H), 1.79 (m,5H); LC/MS (ESI+): 474.0 (M+H).

Assignment of Absolute Stereochemistry for Example 16b. X-Ray Data forExample 17

Enantiomerically pure Example 16b was derivatized to Example 17 as shownin Scheme 5 and in the previous section. The absolute stereochemistrywas established by x-ray crystallography, as described below. Based onthe heavy-atom effect (cf also Mudianta, I. W. Katavic, P. L.; Lambert,L. K. et al. Tetrahedron 2010, 66, 2752 for other examples) of thebromine atom, the absolute stereochemistry of Examples 17 and 16b wasassigned via anomalous dispersion x-ray crystallography to be (3R,4R).The ORTEP diagram is shown in FIG. 3.

X-Ray Crystallography Experimental Procedure—Example 17.

All reflection intensities were measured using a KM4/Xcalibur (detector:Sapphire3) with enhance graphite-monochromated Mo Kα radiation(λ=0.71073 Å) under the program CrysAlisPro (Version 1.171.33.55, OxfordDiffraction Ltd., 2010). The program CrysAlisPro (Version 1.171.33.55,Oxford Diffraction Ltd., 2010) was used to refine the cell dimensions.Data reduction was done using the program CrysAlisPro (Version1.171.33.55, Oxford Diffraction Ltd., 2010). The structure was solvedwith the program SHELXS-97 (Sheldrick, 2008) and was refined on F² withSHELXL-97 (Sheldrick, 2008). Analytical numeric absorption correctionsbased on a multifaceted crystal model were applied using CrysAlisPro(Version 1.171.33.55, Oxford Diffraction Ltd., 2010). The temperature ofthe data collection was controlled using the system Cryojet(manufactured by Oxford Instruments). The H-atoms (except for the Hatoms located on N2n and O3n, n=A, B) were placed at calculatedpositions using the instructions AFIX 13, AFIX 23, AFIX 43 or AFIX 137with isotropic displacement parameters having values 1.2 or 1.5 timesUeq of the attached C atoms. The H-atoms located on N2n and O3n (n=A, B)were found from difference Fourier maps, and their positions wererestrained so that the N—H and O—H distances refine to 0.88(3) and0.84(3) A, respectively.

Data were collected at 110(2) K after the crystal has been flash-cooled.The structure of the compound C₃₄H₃₄BrN₅O₅ was solved and refined in thenon-centrosymmetric space group P1 with Z′=2 (i.e., there are twocrystallographically independent molecules in the asymmetric unit). Thestructure of Example 17 is ordered. The absolute configuration wasestablished by anomalous-dispersion effects in diffraction measurementson the crystal, and the model has chirality R on C11n and C12n (n=A, B).The two chiral centers are found in the trans conformation. The Flackparameter refines to −0.019(5) (note: a value of 0 for this parameterindicated that the absolute structure given by the structure refinementhas been correctly assigned).

The final refinement against F² was acceptable. The R factor [F²>2σ(F²)]is about 0.044. The final difference Fourier map was relatively flat,and the residual peaks are no larger than 0.67 e Å⁻³. The residual peaksQ1 (0.67 e Å⁻³) and Q3 (0.43 e Å⁻³) seems to be found at sites, whichmight be potential acceptors in N—H′″A (A=acceptor). However, thesepeaks are too small to be lattice water molecules. C₃₄H₃₄BrN₅O5,Fw=672.57, yellow rod, 0.40×0.15×0.10 mm³, triclinic, P1 (no. 1),a=7.32713(12), b=11.12781(17), c=19.6664(3) Å, α=106.2111(14),β=91.7015(13), γ=93.2437(12)°, V=1535.54(4) Å³, Z=2, D_(x)=1.455 g cm³,μ=1.390 mm⁻¹, abs. corr. range: 0.631-0.905. 37796 Reflections weremeasured up to a resolution of (sin θ/λ)_(max)=0.62 Å⁻¹. 12019Reflections were unique (R_(int)=0.0380), of which 10778 were observed[I>2σ(I)]. 827 Parameters were refined with 7 restraints. R1/wR2[I>2σ(I)]: 0.0442/0.1187. R1/wR2 [all refl.]: 0.0486/0.1202. S=1.092.Residual electron density found between −0.47 and 0.68 e Å⁻³.

Biological Data

Experimental procedures for determination of inhibitory activity (IC₅₀)against ALK autophosphorylation in enzyme assay and cellular assay wereperformed as described in U.S. Pat. No. 8,471,005 and in Cheng M., QuailM. R., Gingrich D. E., et al. Mol Cancer Ther 2012; 11, 670-679. Theinhibitory activities for the exemplified compounds are given in Table1.

TABLE 1 ALK Inhibitory activity for Examples 1-18. ALK IC50 ALK CellIC50 Example (nM) (nM) 1 3.33 50 2 8.23 80 3 9.51 60 4 13.98 250 5 5.66250 6 11.13 120 7 9.48 130 8 12.76 9 28.16 10  25.19 11  34.03 12  13.74200 13  9.81 250 14  3.00 60 14a 4.12 40 14b 2.23 40 15a 6.53 60 15b9.61 200 16  5.00 50 16a 3.76 100 16b 3.52 120 17  18  3.00 150

Rat Pharmacokinetic (Pk) Studies.

Experimental protocols for rat PK determinations followed procedurespreviously described (Ott, G. R.; Wells, G. J.; Thieu, T. V.; Quail, M.R. et al. J Med. Chem. 2011, 54, 6328-6341). Examples 14 (racemic, cis)and 16 (racemic, trans) were identified as having acceptable rat PKproperties. See Table 2 below.

TABLE 2 Sprague-Dawley Rat Pharmacokinetic Parameters. PK prameters 1416 i.v. dose (mg/kg) 1 1 t_(1/2) (h)  2.5 ± 0.6  3.2 ± 0.6 AUC_(0-∞)(ng*h/mL) 371 ± 6  459 ± 72 Vd (L/kg) 9.9 ± 2  10.3 ± 1.3 CL (mL/min/kg)46 ± 1 39 ± 7 p.o. dose (mg/kg) 5 5 C_(max) (ng/mL)  85 ± 15  97 ± 10t_(max) (h)  6 ± 0   4 ± 1.2 AUC_(0-∞) (ng*h/mL) 479 ± 83 708 ± 63 F %26 ± 4 31 ± 3The rat PK parameters for the two diastereomeric racemates Example 14and Example 16 were comparable.

In Vivo Studies General Experimental Procedures:

In vivo PK/PD and tumor growth inhibition studies in rodents wereperformed according to the protocols described below, previouslypublished in: Cheng M., Quail M. R., Gingrich D. E., et al. Mol CancerTher 2012; 11, 670-679 and Ott, G. R.; Wells, G. J.; Thieu, T. V.;Quail, M. R. et al. J. Med. Chem. 2011, 54, 6328-6341.

Cell Lines

The NPM-ALK-positive ALCL cell lines, Karpas-299 and Sup-M2, werepurchased from Deutsche Sammlung von Mikroorganismen und ZellkulturenGmbH.

Animals

Severe combined immunodeficient (SCID)/beige or nu/nu mice (6- to8-week-old female) were maintained 5 per cage in microisolator units ona standard laboratory diet (Teklad Labchow). Animals were housed underhumidity- and temperature-controlled conditions and the light/dark cyclewas set at 12-hour intervals, maintained under specified andopportunistic pathogen-free conditions. Mice were quarantined at least 1week before experimental manipulation. All animal studies were conductedunder protocol approved by the Institutional Animal Care and UseCommittee of Cephalon, Inc. or by University of Turin Ethical Committee.

Immunoblot Analysis

Immunoblotting of phospho- and total ALK as well as the downstreamtargets was carried out according to the protocols provided by theantibody suppliers. In brief, after treatment, cells were lysed in FRAKlysis buffer [10 mmol/L Tris, pH 7.5, 1% Triton X-100, 50 mmol/L sodiumchloride, 20 mmol/L sodium fluoride, 2 mmol/L sodium pyrophosphate, 0.1%BSA, plus freshly prepared 1 mmol/L activated sodium vanadate, 1 mmol/LDTT, and 1 mmol/L phenylmethylsulfonylfluoride, protease inhibitorscocktail III (1:100 dilution, catalog no. 539134; Calbiochem)]. Afterbrief sonication, the lysates were cleared by centrifugation, mixed withsample buffer, and subjected to SDS-PAGE. Following transfer tomembranes, the membranes were blotted with individual primary andsecondary antibodies, washed in TBS/0.2% Tween, and protein bandsvisualized with Enhanced Chemiluminescence. The individual bands ofphospho- and total NPM-ALK were scanned and quantified with the Gel-ProAnalyzer software (Media Cybernetics, Inc.).

PK/PD Studies

Exponentially growing cells were implanted subcutaneously to the leftflank of each mouse. The mice were monitored and when the tumorxenograft volumes reached approximately 300 to 500 mm3, mice received asingle oral administration of either vehicle PEG-400 or the compound ofinterest (e.g., Examples 14, 16, 16a, and/or 16b) formulated in vehicle.At indicated time points postdosing, the mice were sacrificed, the bloodwas collected and centrifuged, and the plasma was collected. The tumorswere excised and disrupted with a hand-held tissue blender in completedFRAK lysis buffer without Triton X-100. After brief sonication, thelysates were cleared by centrifugation, mixed with sample buffer, andsubjected to SDS-PAGE for ALK immunoblotting as described above. Theindividual bands of phospho- and total NPM-ALK were scanned andquantified with the Gel Pro Analyzer software (Media Cybernetics, Inc.).The relative NPM-ALK tyrosine phosphorylation (phospho-NPMALK/NPM-ALKratio) of each sample at indicated time points was then calculated, withthe average value of vehicle-treated sample(s) as 100. The compoundlevels in plasma and tumor lysates were measured by liquidchromatography/tandem mass spectrometry (LC/MS-MS).

PK/PD Results and Discussion

Racemic Examples 14 (cis relative stereochemistry) and 16 (transrelative stereochemistry) showed favorable and comparable in vitroprofiles and rat PK properties (Table 2). Therefore, to evaluate theirability to inhibit the autophosphorylation of NPM-ALK in vivo, bothracemic compounds were advanced into single dose PK/PD experiments insubcutaneous ALK-dependent Sup-M2 xenografts in Scid mice (FIGS. 1A and1B, respectively). In these experiments, racemic Example 14 exhibitedmodest inhibition of ALK phosphorylation when dosed orally at 30 mg/kg,with <75% inhibition relative to the vehicle-only treated animals, atany time point up to 24 h (FIG. 1A). The separated, single enantiomersof Example 14, Examples 14a and 14b both showed comparably modestinhibition when dosed at 30 mg/kg in independent PK/PD experiments (datanot shown).

To contrast, racemic Example 16 (trans) effected 75-87% inhibition ofALK phosphoryation up to 12 h after an equivalent dose, albeit followedby recovery of the signal at the 24 h time point (FIG. 1B). Thispositive result was unpredictable in view of the comparable in vitrodata between Examples 14 and 16. Furthermore, the separated singleenantiomers Example 16a and Example 16b gave different in vivo PK/PDresponses when tested independently, which is an unpredicted resultbased on their comparable in vitro activity profiles. Specifically,Example 16a has an enzyme IC50 of 3.76 nM and a cell IC50 of 100 nM,whereas Example 16b has an enzyme IC50 of 3.52 nM and a cell IC50 of 120nM. (See Table 1) Despite the comparable in vitro data, singleenantiomer Example 16a had only a modest impact on the ALKautophosphorylation: 40-60% inhibition over 16 h post single p.o. doseof 30 mg/kg (FIG. 1C), whereas single enantiomer Example 16b inhibited80-100% of the signal up to 16 h, under the same experimental conditions(FIG. 1D). Plasma and tumor levels achieved with Example 16b (FIG. 1F)at 6 h and 16 h post dose were also unexpectedly higher (in the 4-10fold range) than the plasma and tumor levels achieved with Example 16aat the same time-points (FIG. 1E). In view of the discovery of thesestereoselective in vivo PK/PD responses, the preferred single enantiomerExample 16b was advanced in antitumor efficacy studies, which aredescribed below.

Antitumor Efficacy Studies

Tumor-bearing mice were randomized into different treatment groups (8-10mice per group) and administered orally either vehicle (PEG-400, ordH2O) or Example 16b formulated in vehicle at indicated doses (expressedas mg/kg equivalents of free base) and with indicated dosing frequency,with 100 mL per dosing volume. The length (L) and width (W) of eachtumor was measured with a Vemier caliper and the mouse body weight wasdetermined every 2 to 3 days. The tumor volumes were then calculatedwith the formula of 0.5236*L*W*(L+W)/2. Statistical analyses of tumorvolumes and mouse body weight were carried out using the Mann-Whitneyrank sum test. Plasma and tumor samples were obtained at 2 hourspost-final dose, and the compound levels in plasma and tumor lysateswere measured by LC/MS-MS.

Based upon the robust PK/PD response, (FIG. 1D), the single enantiomerExample 16b was selected for anti-tumor efficacy studies in Sup-M2xenografts in Scid mice. Complete tumor regressions were observed upontreatment with either 30 mg/kg or 55 mg/kg of Example 16b b.i.d., p.o.doses for 10 days (FIG. 2). No remaining tumor could be observed after10 days of dosing. Both dosing regimens were well tolerated, with nobody weight loss or other overt toxicity being detected. Such positiveresults in the in vivo PK/PD and Antitumor Efficacy studies with thesingle enantiomer Example 16b were not predictable in view of the invitro or in vivo data for Example 14, Example 16 or Example 16a.

1. A compound that is a single enantiomer selected from the groupconsisting of:

and/or a salt thereof.
 2. The single enantiomer according to claim 1that is(3R,4R)-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol,and/or a salt thereof.
 3. A compound selected from the group consistingof:(±)-3,4-cis-1-(2-Methoxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol;(±)-3,4-cis-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol;(±)-2-(3,4-trans-3-Fluoro-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol;(±)-3,4-cis-4-{3-Methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol;(±)-[4-(3,4-trans-3-Fluoro-piperidin-4-yl)-2-methoxy-phenyl]-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-yl]-amine;(±)-2-(3,4-trans-3-fluoro-4-{3-methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol;(±)-2-(3,3-Difluoro-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol;(±)-1-(3,4-trans-3-Fluoro-4-{3-methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-2-methyl-propan-2-ol;(±)-2-(3,3-Difluoro-4-{3-methoxy-4-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol;(±)-[4-(3,3-Difluoro-piperidin-4-yl)-2-methoxy-phenyl]-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-yl]-amine;(±)-[4-(3,4-trans-3-Fluoro-piperidin-4-yl)-2-methoxy-phenyl]-[7-(2-methoxy-pyridin-3-yl)-pyrrolo[2,1-f][1,2,4]triazin-2-yl]-amine;(3R,4S)-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol;(3S,4R)-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol;2-((3S,4S)-3-Fluoro-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol;2-((3R,4R)-3-Fluoro-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol;(3S,4S)-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol;(3R,4R)-1-(2-Hydroxy-ethyl)-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-3-ol;4-Bromo-benzoic acid2-((3R,4R)-3-hydroxy-4-{3-methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethylester;2-(4-{3-Methoxy-4-[7-(2-methoxy-phenyl)-pyrrolo[2,1-f][1,2,4]triazin-2-ylamino]-phenyl}-piperidin-1-yl)-ethanol;and a salt thereof.
 4. A pharmaceutical composition comprising acompound according to claim 1, and/or a salt thereof, and apharmaceutically acceptable excipient.
 5. A method of inhibiting ALKkinase in a subject in need thereof, comprising administering to thesubject a therapeutically effective amount of a compound according toclaim 1, and/or a salt thereof.
 6. A method of treating a ALK-positivedisease/disorder in a patient in need thereof, comprising administeringto the subject a therapeutically effective amount of a compoundaccording to claim 1, and/or a salt thereof.
 7. A method of treatingproliferative disease in a patient in need thereof, comprisingadministering to the subject a therapeutically effective amount of acompound according to claim 1, and/or a salt thereof.
 8. Apharmaceutical composition comprising a compound according to claim 3,and/or a salt thereof, and a pharmaceutically acceptable excipient.
 9. Amethod of inhibiting ALK kinase in a subject in need thereof, comprisingadministering to the subject a therapeutically effective amount of acompound according to claim 3, and/or a salt thereof.
 10. A method oftreating a ALK-positive disease/disorder in a patient in need thereof,comprising administering to the subject a therapeutically effectiveamount of a compound according to claim 3, and/or a salt thereof.
 11. Amethod of treating proliferative disease in a patient in need thereof,comprising administering to the subject a therapeutically effectiveamount of a compound according to claim 3, and/or a salt thereof.